47169 and 33935, novel human glycosyl transferases and uses therefor

Abstract

The invention provides isolated nucleic acids molecules, designated 47169 and 33935 nucleic acid molecules, which encode novel glycosyl transferases. The invention also provides antisense nucleic acid molecules, recombinant expression vectors containing 47169 and 33935 nucleic acid molecules, host cells into which the expression vectors have been introduced, and non-human transgenic animals in which a 47169 or 33935 gene has been introduced or disrupted. The invention still further provides isolated 47169 and 33935 proteins, fusion proteins, antigenic peptides and anti-47169 and anti-33935 antibodies. Diagnostic methods utilizing compositions of the invention are also provided.

1 - 35 . (canceled)  
     
     
         36 . A method for identifying a candidate compound capable of modulating cellular proliferation, the method comprising: 
 a) contacting a test compound with a sample comprising a polypeptide selected from the group consisting of: 
 i) a polypeptide which is at least 95% identical to the amino acid sequence of SEQ ID NO:2, wherein the polypeptide has glycosyl transferase activity;  
 ii) a polypeptide which is encoded by a nucleic acid molecule which is at least 95% identical to the nucleotide sequence of SEQ ID NO:1 or 3, wherein the polypeptide has glycosyl transferase activity; and  
 iii) a polypeptide comprising a fragment of at least 100 contiguous amino acids of SEQ ID NO:2, wherein the polypeptide has glycosyl transferase activity;  
   b) assaying the ability of the test compound to modulate the activity of the polypeptide;    c) contacting a cell which expresses the polypeptide with the compound identified in b); and    d) determining the effect of the compound on cellular proliferation;    thereby identifying a compound capable of modulating cellular proliferation.    
     
     
         37 . The method of  claim 36 , wherein the glycosyl transferase activity is the ability to transfer an N-acetylgalactosamine moiety from uridine diphosphate to a hydroxyl moiety of a serine or threonine residue of a protein.  
     
     
         38 . The method of  claim 36 , wherein the sample comprises the polypeptide, a membrane-bound form of an isolated polypeptide or a cell expressing the polypeptide.  
     
     
         39 . The method of  claim 38 , wherein the cell is selected from the group consisting of a colon cell, an ovarian cell, a breast cell, a lung cell and a liver cell.  
     
     
         40 . The method of  claim 36 , wherein the compound is a small molecule, a peptide, or an antibody.  
     
     
         41 . The method of  claim 36 , wherein the polypeptide further comprises heterologous sequences.  
     
     
         42 . A method for identifying a candidate compound capable of modulating cellular proliferation, the method comprising: 
 a) contacting a test compound with a sample comprising a polypeptide selected from the group consisting of: 
 i) a polypeptide comprising the amino acid sequence of SEQ ID NO:2; and  
 ii) a polypeptide which is encoded by the nucleotide sequence of SEQ ID NO:1 or 3;  
   b) assaying the ability of the test compound to modulate the activity of the polypeptide;    c) contacting a cell which expresses the polypeptide with the compound identified in b); and    d) determining the effect of the compound on cellular proliferation;    thereby identifying a compound capable of modulating cellular proliferation.    
     
     
         43 . The method of  claim 42 , wherein the sample comprises the polypeptide, a membrane-bound form of an isolated polypeptide or a cell expressing the polypeptide.  
     
     
         44 . The method of  claim 43 , wherein the cell is selected from the group consisting of a colon cell, an ovarian cell, a breast cell, a lung cell and a liver cell.  
     
     
         45 . The method of  claim 42 , wherein the compound is a small molecule, a peptide, or an antibody.  
     
     
         46 . The method of  claim 42 , wherein the polypeptide further comprises heterologous sequences.  
     
     
         47 . A method for identifying a candidate compound capable of modulating cellular proliferation, the method comprising: 
 a) combining a test compound with a sample comprising a polypeptide selected from the group consisting of: 
 i) a polypeptide which is at least 95% identical to the amino acid sequence of SEQ ID NO:2, wherein the polypeptide has glycosyl transferase activity;  
 ii) a polypeptide which is encoded by a nucleic acid molecule which is at least 95% identical to the nucleotide sequence of SEQ ID NO:1 or 3, wherein the polypeptide has glycosyl transferase activity; and  
 iii) a polypeptide comprising a fragment of at least 100 contiguous amino acids of SEQ ID NO:2, wherein the polypeptide has glycosyl transferase activity;  
 under conditions suitable for binding;  
   b) selecting a compound capable of binding to the polypeptide;    c) contacting a cell which expresses the polypeptide with the compound identified in b); and    d) determining the effect of the compound on cellular proliferation;    thereby identifying a compound capable of modulating cellular proliferation.    
     
     
         48 . The method of  claim 47 , wherein the glycosyl transferase activity is the ability to transfer an N-acetylgalactosamine moiety from uridine diphosphate to a hydroxyl moiety of a serine or threonine residue of a protein.  
     
     
         49 . The method of  claim 47 , wherein the sample comprises the polypeptide, a membrane-bound form of an isolated polypeptide or a cell expressing the polypeptide.  
     
     
         50 . The method of  claim 49 , wherein the cell is selected from the group consisting of a colon cell, an ovarian cell, a breast cell, a lung cell and a liver cell.  
     
     
         51 . The method of  claim 47 , wherein the compound is a small molecule, a peptide, or an antibody.  
     
     
         52 . The method of  claim 47 , wherein the polypeptide further comprises heterologous sequences.  
     
     
         53 . The method of  claim 47 , wherein the binding of the test compound to the polypeptide is determined by a method selected from the group consisting of: 
 a) direct detecting of test compound/polypeptide binding;    b) a competition binding assay;    c) an immunoassay;    d) a yeast two-hybrid assay; and    e) an assay detecting the formation of a covalent bond between a carbohydrate (i.e., saccharide) moiety and a hydroxyl or amino moiety of a protein.    
     
     
         54 . A method for identifying a candidate compound capable of modulating cellular proliferation, the method comprising: 
 a) combining a test compound with a sample comprising a polypeptide selected from the group consisting of: 
 i) a polypeptide comprising the amino acid sequence of SEQ ID NO:2; and  
 ii) a polypeptide which is encoded by the nucleotide sequence of SEQ ID NO:1 or 3;  
 under conditions suitable for binding;  
   b) selecting a compound capable of binding to the polypeptide;    c) contacting a cell which expresses the polypeptide with the compound identified in b); and    d) determining the effect of the compound on cellular proliferation;    thereby identifying a compound capable of modulating cellular proliferation.    
     
     
         55 . The method of  claim 54 , wherein the sample comprises the polypeptide, a membrane-bound form of an isolated polypeptide or a cell expressing the polypeptide.  
     
     
         56 . The method of  claim 55 , wherein the cell is selected from the group consisting of a colon cell, an ovarian cell, a breast cell, a lung cell and a liver cell.  
     
     
         57 . The method of  claim 54 , wherein the compound is a small molecule, a peptide, or an antibody.  
     
     
         58 . The method of  claim 54 , wherein the polypeptide further comprises heterologous sequences.  
     
     
         59 . The method of  claim 54 , wherein the binding of the test compound to the polypeptide is determined by a method selected from the group consisting of: 
 a) direct detecting of test compound/polypeptide binding;    b) a competition binding assay;    c) an immunoassay;    d) a yeast two-hybrid assay; and    e) an assay detecting the formation of a covalent bond between a carbohydrate (i.e., saccharide) moiety and a hydroxyl or amino moiety of a protein.